caa a22 4500 606212078 CHVBK 20210128101041.0 cr unu---uuuuu 210128e20150601xx s 000 0 eng 10.1007/s10295-015-1608-7 doi (NATIONALLICENCE)springer-10.1007/s10295-015-1608-7 Overexpression and characterization of a Ca2+ activated thermostable β-glucosidase with high ginsenoside Rb1 to ginsenoside 20(S)-Rg3 bioconversion productivity [Elektronische Daten] [Jingcong Xie, Dongxia Zhao, Linguo Zhao, Jianjun Pei, Wei Xiao, Gang Ding, Zhenzhong Wang] The thermostable β-glucosidase gene from Thermotoga petrophila DSM 13995 was cloned and overexpressed in Escherichia coli. The activity of the recombinant β-glucosidase was 21U/mL in the LB medium. Recombinant β-glucosidase was purified, and its molecular weight was approximately 81kDa. The optimal activity was at pH 5.0 and 90°C, and the thermostability of the enzyme was improved by Ca2+. The β-glucosidase had high selectivity for cleaving the outer and inner glucopyranosyl moieties at the C-20 carbon of ginsenoside Rb1, which produced the pharmacologically active minor ginsenoside 20(S)-Rg3. In a reaction at 90°C and pH 5.0, 10g/L of ginsenoside Rb1 was transformed into 6.93g/L of Rg3 within 90min, with a corresponding molar conversion of 97.9%, and Rg3 productivity of 4620mg/L/h. This study is the first report of a GH3-family enzyme that used Ca2+ to improve its thermostability, and it is the first report on the high substrate concentration bioconversion of ginsenoside Rb1 to ginsenoside 20(S)-Rg3 by using thermostable β-glucosidase under high temperature. Society for Industrial Microbiology and Biotechnology, 2015 Thermotoga petrophila nationallicence Recombinant enzyme nationallicence Enzymatic characterization nationallicence Biotransformation nationallicence Ginsenoside 20( S )-Rg3 preparation nationallicence Xie Jingcong College of Chemical Engineering, Nanjing Forestry University, 159 Long Pan Road, 210037, Nanjing, China aut Zhao Dongxia College of Chemical Engineering, Nanjing Forestry University, 159 Long Pan Road, 210037, Nanjing, China aut Zhao Linguo College of Chemical Engineering, Nanjing Forestry University, 159 Long Pan Road, 210037, Nanjing, China aut Pei Jianjun College of Chemical Engineering, Nanjing Forestry University, 159 Long Pan Road, 210037, Nanjing, China aut Xiao Wei Jiangsu Kanion Pharmaceutical Co., Ltd., 58 Haichang South Road, 222001, Lianyungang, China aut Ding Gang Jiangsu Kanion Pharmaceutical Co., Ltd., 58 Haichang South Road, 222001, Lianyungang, China aut Wang Zhenzhong Jiangsu Kanion Pharmaceutical Co., Ltd., 58 Haichang South Road, 222001, Lianyungang, China aut Journal of Industrial Microbiology & Biotechnology Springer Berlin Heidelberg 42/6(2015-06-01), 839-850 1367-5435 42:6<839 2015 42 10295 https://doi.org/10.1007/s10295-015-1608-7 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s10295-015-1608-7 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Xie Jingcong College of Chemical Engineering, Nanjing Forestry University, 159 Long Pan Road, 210037, Nanjing, China aut NATIONALLICENCE 700 1- Zhao Dongxia College of Chemical Engineering, Nanjing Forestry University, 159 Long Pan Road, 210037, Nanjing, China aut NATIONALLICENCE 700 1- Zhao Linguo College of Chemical Engineering, Nanjing Forestry University, 159 Long Pan Road, 210037, Nanjing, China aut NATIONALLICENCE 700 1- Pei Jianjun College of Chemical Engineering, Nanjing Forestry University, 159 Long Pan Road, 210037, Nanjing, China aut NATIONALLICENCE 700 1- Xiao Wei Jiangsu Kanion Pharmaceutical Co., Ltd., 58 Haichang South Road, 222001, Lianyungang, China aut NATIONALLICENCE 700 1- Ding Gang Jiangsu Kanion Pharmaceutical Co., Ltd., 58 Haichang South Road, 222001, Lianyungang, China aut NATIONALLICENCE 700 1- Wang Zhenzhong Jiangsu Kanion Pharmaceutical Co., Ltd., 58 Haichang South Road, 222001, Lianyungang, China aut NATIONALLICENCE 773 0- Journal of Industrial Microbiology & Biotechnology Springer Berlin Heidelberg 42/6(2015-06-01), 839-850 1367-5435 42:6<839 2015 42 10295